Identification of a new dysfunctional platelet P2Y12 receptor variant associated with bleeding diathesis.

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Date publication

février 2015

Journal

Blood

Auteurs

Membres identifiés du Cancéropôle Est :
Dr GACHET Christian


Tous les auteurs :
Lecchi A, Razzari C, Paoletta S, Dupuis A, Nakamura L, Ohlmann P, Gachet C, Jacobson KA, Zieger B, Cattaneo M

Résumé

Defects of the platelet P2Y12 receptor (P2Y12R) for adenosine diphosphate (ADP) are associated with increased bleeding risk. The study of molecular abnormalities associated with inherited qualitative defects of the P2Y12R protein is useful to unravel structure-function relationships of the receptor. We describe the case of 2 brothers, sons of first cousins, with lifelong history of abnormal bleeding, associated with dysfunctional P2Y12R and a previously undescribed missense mutation in the encoding gene. ADP (4-20 µM)-induced aggregation of patients' platelets was markedly reduced and rapidly reversible. Other agonists induced borderline-normal aggregation. Inhibition of vasodilator-stimulated phosphoprotein phosphorylation and prostaglandin E1-induced increase in cyclic adenosine monophosphate (cAMP) by ADP was impaired, whereas inhibition of cAMP increase by epinephrine was normal. [(3)H]PSB-0413, a selective P2Y12R antagonist, bound to a normal number of binding sites; however, its affinity, and that of the agonists ADP and 2-methylthio-adenosine-5'-diphosphate, was reduced. Patients' DNA showed a homozygous c.847T>A substitution that changed the codon for His-187 to Gln (p.His187Gln). Crystallographic data and molecular modeling studies indicated that His187 in transmembrane 5 is important for agonist and nucleotide antagonist binding and located in a region undergoing conformational changes. These studies delineate a region of P2Y12R required for normal function after ADP binding.

Mots clés

Adenosine Diphosphate, metabolism, Adenosine Monophosphate, analogs & derivatives, Blood Platelet Disorders, genetics, Blood Platelets, metabolism, Cyclic AMP, metabolism, Hemorrhagic Disorders, genetics, Humans, Male, Middle Aged, Molecular Docking Simulation, Pedigree, Platelet Aggregation, Point Mutation, Receptors, Purinergic P2Y12, genetics, Thionucleosides, metabolism

Référence

Blood. 2015 Feb;125(6):1006-13